...advantage to using the CRISPR/Cas system for genome engineering is the fact that Cas9 can be easily programmed...smegmatis (Yan et al., 2020). NHEJ can also be engaged by variants of the canonical Cas9 approach. A pair...UNC Chapel Hill.   References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM (2009) Limiting the Persistence...

... evaluate degradation of ENL. Cell lines were engineered to express ENL-FKBP12F36V and endogenous ENL ...evaluate degradation of MELK. Cell lines were engineered to express FKBP12F36V-MELK and endogenous MELK... evaluate degradation of YY1. Cell lines were engineered to express YY1-FKBP12F36V using CRISPR/Cas9-mediated...

... is a postdoc at Harvard University where she engineers genetically encoded voltage indicators. Tian developed...mutant sensors expressed in a mammalian cell line engineered to be electrically excitable. The mutants are.... Tian notes that this platform can be used to engineer many types of biosensors.   This screening method...

...injected with an immunogen. This might be the full-length protein that you want to generate antibodies against...Otherwise, you might experience variability in the strength of staining and background noise. If you do use...control across experiments to be aware of signal strength. In general, polyclonal antibodies produce a higher...

...excitation wavelength. A fluorophore is less likely to bleach when it is excited off the peak wavelength, since... excitation powers are often used to reduce the length of the experiments. As the photobleaching rate ...

...with him.  He is also available for speaking engagements as are many of the coaches I have mentioned. ...Consulting works remotely, will travel for speaking engagements and has many areas of specialty from leadership...but the website looks interesting and it is in English and Spanish. She is based in Spain. The bottom ...

...high-complexity reactions? The researchers over at New England Biolabs® thought otherwise! They suspected a data-driven... Golden Gate Assemblies. Image courtesy of New England Biolabs.   DAD Tools The three DAD tools used...Temperature Incubation time/cycling Overhang length T4 DNA ligase 25 °C 1 hour 3-base or ...

...particularly hard. There are millions of them. The real challenge is finding relevant papers. The latest installment...literature! PubMed PubMed is the classic search engine for finding papers. It’s been around since 1996...Zuckerberg Initiative. Meta is not a traditional search engine. Instead, you create your own research feed by ...

...CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability ...tool to the CRISPR tool box.  Nanoblades are engineered murine leukemia virus (MLV)-like particles loaded...pronucleus of the zygote which is both technically challenging and can negatively impact zygote viability. Nanoblades...

...its ground state by emission of a photon with wavelength specific to the acceptor In order for FRET to...quantify mechanical tension (a molecular “spring”) (Meng et al., 2008), and monitor neuronal activity (voltage...Nature 388.6645 (1997): 882. PubMed PMID: 9278050. 3. Meng, Fanjie, et al. "A fluorescence energy transfer‐...

...Note: Cpf1 is also called Cas12a. In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant...editing. LbCpf1 and AsCpf1 are 3.7 kb and 3.9 kb in length, respectively, larger than SaCas9 but smaller than...deletions at the 3’ end of the target sequence. Cpf1’s strength may lie in its low off-target editing rates, determined...

...fiber optics needed for optogenetics. First, the length of the fiber optic for the implant depends on the...started, you will need a laser of the appropriate wavelength to stimulate your opsin. Each opsin has an excitation...fiber optic patch cable must be the appropriate length for your specific behavioral apparatus or test ...

...particular protein DNA must first be transcribed into messenger RNA (mRNA). mRNA can then be translated at the... however, are expensive and very technically challenging to use. Alyssa D. Cecchetelli is a Scientist... expression in Escherichia coli: advances and challenges Production of recombinant proteins in plant root...

...motivates you!), know their unique skills and strengths (you will likely have reports with different levels.... Personally I found this to be particularly challenging. As PhD students and postdocs we are used to ...was a manager! Second, my direct report was not engaged because I only left him/her with small tasks and...

... to lab-adapted bacteria because it has been challenging to introduce CRISPRi machinery into diverse bacteria...antibiotics are often essential genes which creates a challenge when studying them since their complete knockdown...about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi, and CRISPRa...

...scientists work to invent novel techniques or to engineer improved tools for familiar applications. But ...variants of MagLOV will be possible with other engineering approaches and that magnetosensitive tools can...biophysics, as well as inventors and biomedical engineers, join efforts to explore the mechanisms and develop...

... they will emit fluorescent light at similar wavelengths (See this handy emission spectra chart to find...the two of them because they emit at a similar wavelength. However, if your flow cytometer reads them out...Compensating between your fluorescent markers can be challenging! But we hope this guide can help you get started...

...kDa). This system also requires less genetic engineering than some of the other systems reviewed here....PROTAC) was developed for HaloTag. PROTACs are engineered molecules which target E3 ligases to specific...that it requires an additional component to be engineered or expressed beyond the tag and small molecule...

...postings. Also, if you work in industry for any length of time, you’ll know more people in industry, which...in-depth discussion about your project might be challenging. In an academic environment, surrounded by lab...AstraZeneca, Pfizer, Novartis, Biogen, Amgen, New England Biolabs, Regeneron, and Genentech, among others...

...Adenoviral vector DNA for accurate genome editing with engineered nucleases. Nat Methods 11:1051–1057 . https:/...Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering of oncogenic chromosomal rearrangements with ...Tran K, Li J, Yin H, Anderson DG, Sontheimer EJ, Weng Z, Gao G, Xue W (2015) Adenovirus-Mediated Somatic...